2.1. Łagodzenie stresu (ID 938)

The claimed effect, which is proposed for further assessment, is “significant improvement of stress- induced psychological and gastrointestinal symptoms like anxiety, anger-hostility, depressive symptoms, nausea and abdominal pain”. The proposed target population is healthy adults experiencing moderate stress or anxiety.
The Panel assumes that the claimed effect refers to alleviation of psychological stress. The Panel considers that alleviation of psychological stress is a beneficial physiological effect.

2.2. Utrzymanie równowagi prozdrowotnej flory bakteryjnej w celu zwiększenia naturalnej odporności organizmu (ID 2942)

The claimed effect, which is proposed for further assessment, is “maintains the balance of healthy microbiota that helps to strengthen the natural defence”. The proposed target population is the general population.
The Panel notes that it is not possible to define the exact number of the different microbial groups which constitute a balanced healthy microbiota.
The references provided in relation to this claim included reports on identification and characterisation aspects of the bacterial strains (Naser et al., 2006), a review on anti-inflammatory activity of different bacterial strains (Mengheri, 2008), an in vitro study on the properties of the bacterial strains (i.e. adhesion, competition against enteropathogens and modulation of IL-8 production (Candela et al., 2008)), an animal study investigating the effect of the combination of the strains which is the subject of the claim on an experimental model of induced colitis in mice (Roselli et al., 2009), and a human intervention study investigating the effect of a combination of the strains containing also fructo-oligosaccharides on the composition of gut microbiota and metabolic profiles (Vitali et al., 2010). An abstract from a meeting was also provided; the abstract reported on a human study conducted with the combination of the strains that is the subject of the claim on lymphocyte subsets from elderly subjects (Finamore, 2011). The Panel notes that from the information provided it was not possible to establish the specific effect which is the subject of the claim.
The Panel considers that the claimed effect is general and non-specific, and does not refer to any specific health claim as required by Regulation (EC) No 1924/2006.

3.1. Łagodzenie stresu (ID 938)

Among the references provided in relation to the claim were human, animal and in vitro studies which were unrelated to the combination of L. helveticus CNCM I-1722 and B. longum CNCM I-3470 (Estrada et al., 2001; Firmesse et al., 2008; Haskey and Dahl, 2006; Johnson-Henry et al., 2007; Wine et al., 2009). The Panel considers that no conclusions can be drawn from these studies for the scientific substantiation of the claim.
Three human intervention studies on a combination of L. helveticus CNCM I-1722 and B. longum CNCM I-3470 (Diop et al., 2008; Messaoudi et al., 2010; 2011) were provided for the scientific substantiation of the claim.
Diop et al. (2008) in a double-blind, placebo-controlled, randomised, parallel study assessed the effect of a combination of L. helveticus CNCM I-1722 and B. longum CNCM I-3470 on symptoms claimed to be related to stress. Adult volunteers aged 18-60 years with at least two symptoms (anxiety, nervousness, irritability, sleeping problems, gastro-intestinal disturbances) perceived as induced by stress during the preceding month, but not receiving any medical treatment for stress-induced symptoms, were eligible for the study. Subjects randomly received a combination of the two bacterial strains (3 x 109 CFU per sachet, L. helveticus CNCM I-1722 and B. longum CNCM I-3470 in a ratio of 9:1, one sachet daily) or placebo for three weeks. Participants completed a questionnaire on stress- induced symptoms at the beginning and at the end of the intervention. The questionnaire consisted of 62 items related to symptoms in the following areas: gastro-intestinal, cardiovascular, sleeping, locomotor, physical, psychological, intellectual, spiritual and social. Each symptom was evaluated using a 10-cm visual analogue scale (VAS). A global score for each area was determined as the mean of each item. The changes of the score between the baseline and the end of the study were calculated and compared between the two groups using the unpaired Student’s t test. A total of 75 subjects (54 females, aged 38±11 years) were randomised (37 in the study group and 38 in the placebo group) and 64 finished the study (31 in the intervention group and 33 in the placebo group). The reasons for drop- outs were given. The Panel notes that no information was given about the validation of the questionnaire used for subjective measurements, that the process of randomisation was insufficiently described, that inclusion criteria (at least two symptoms perceived as induced by stress) were not sufficiently defined and justified, and that no correction for multiple testing was performed in the statistical analysis of the results. The Panel considers that no conclusions can be drawn from this study for the scientific substantiation of the claim.
Messaoudi et al. (2010) studied the potential anxiolytic effect of a combination of L. helveticus CNCM I-1722 and B. longum CNCM I-3470 in a double-blind, placebo-controlled, randomised, parallel study. The Hospital Anxiety and Depression Scale (HADS) was used for the enrolment of subjects. Their results had to be ≤20 in the HADS total score, ≤12 in HADS-anxiety subscale and ≤12 in HADS-depression subscale. Sixty-six subjects were randomised to take either the combination of bacterial strains which is the subject of the claim (in the form of sticks, 3 x 109 CFU daily, L. helveticus CNCM I-1722 and B. longum CNCM I-3470 content in a ratio of 90:10) or placebo, identical in taste and appearance, for 30 days. Fifty-five subjects finished the study (26 subjects in the intervention group, 29 subjects in the control group, mean age 42.4 years in the intervention group and 43.2 years in the control group). At the beginning and at the end of the intervention the participants completed a set of questionnaires: the Hopkins Symptom Checklist (HSCL-90), the HADS, the Perceived Stress Scale (PSS) and the coping checklist (CCL). Moreover, 24 h urinary free cortisol was measured. In a separate publication (Messaoudi et al., 2011), a secondary sub-group analysis of the results of the above-described study for 25 subjects (10 in the study group and 15 in the control group) with urinary free cortisol concentrations less than 50 ng/ml at baseline was presented. The Panel notes that this study was designed for measuring anxiety and not changes induced by stress. The Panel considers that no conclusions can be drawn from these studies for the scientific substantiation of the claim.
The submitted studies in animals evaluated the results of administration of the combination of bacterial strains which is the subject of the claim on the conditioned defensive burying test (Messaoudi et al., 2010), on depressive behaviour after experimental myocardial infarction (Arseneault-Breard et al., 2011), and on apoptosis in the limbic system after myocardial infarction (Girard et al., 2009). The in vitro studies measured the effect of the combination of bacterial strains which is the subject of the claim on the proliferation rate of splenocytes and on immunoglobuline production (Easo et al., 2002), their survival in simulated gastro-intestinal conditions (Possemiers et al., 2010), the susceptibility of several bacterial strains affected by chemical stress to different antibiotics (Kheadr, 2006; Kheadr et al., 2007), and the effect of various Lactobacillus and Bifidobacterium strains on cytokine production by human intestinal epithelial cells (Wallace et al., 2003). The Panel considers that in the absence of evidence for an effect on the alleviation of
psychological stress in humans, evidence provided in these animal and in vitro studies cannot be used for the scientific substantiation of a claim on alleviation of psychological stress.
The Panel notes that no human intervention studies were provided from which conclusions could be drawn for the scientific substantiation of the claim.
The Panel concludes that a cause and effect relationship has not been established between the consumption of a combination of L. helveticus CNCM I-1722 and B. longum subsp. longum CNCM I-3470 and alleviation of psychological stress.